Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis
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Patient-derived organoids (PDOs) are ideal ex vivo model systems to study cancer progression and drug resistance mechanisms. Here, we present a protocol for measuring drug efficacy in three-dimensional (3D) high-grade serous ovarian cancer PDO cultures through quantification of cytotoxicity using propidium iodide incorporation in dead cells. We also provide detailed steps to analyze proliferation of PDOs using the Ki67 biomarker. We describe steps for sample processing, immunofluorescent staining, high-throughput confocal imaging, and image-based quantification for 3D cultures. For complete details on the use and execution of this protocol, please refer to Lahtinen et al. (2023).1
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 102683 |
| Tidsskrift | STAR Protocols |
| Vol/bind | 4 |
| Udgave nummer | 4 |
| Antal sider | 17 |
| ISSN | 2666-1667 |
| DOI | |
| Status | Udgivet - 2023 |
Bibliografisk note
Funding Information:
This project received funding from the European Union’s Horizon 2020 Research and Innovation Program under grant agreement no. 965193 (DECIDER). We are grateful to Krister Wennerberg for advice, good discussions, and sharing HGSC organoid culture protocols prior to their publication. We acknowledge the Bioimaging Core Facility for the access and assistance in regards to imaging. We acknowledge Tiina Naumanen Dietrich for expert assistance in image acquisition with ImageXpress HT.ai and image analysis with the MetaXpress software, Ahmad Abdul-Al for careful assistance in maintaining and culturing of PDOs, and Peppi Alho for skillful assistance with fresh tumor processing.
Publisher Copyright:
© 2023 The Author(s)
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